Cardiac Niche Influences the Direct Reprogramming of Canine Fibroblasts into Cardiomyocyte-Like Cells

Figure 4

Expression of cardiac and fibroblast markers in ciCLMs. (a) Examples of intracellular FACS analysis for the isolation of Tnnt2+ cells (green) and isotype controls (red) in ciCLMs after 14 days of differentiation. (b) Example of Western blot analysis after 14 days of differentiation showed the presence of cardiac troponin I as a late cardiac marker protein in ciCLMs derived from both GHMT CF and GHMT SF cells. Densitometry quantification performed with ImageJ software showed that Tnni3 was 3,2-fold more expressed in GHMT CF compared to GTMH SF. Troponin I was absent in CF and SF transduced with α-MHC-GFP vector. (c) Expression of the fibroblast marker HSP47, in transduced cells compared to control cells (transduced with only α-MHC-GFP vector). HSP47 expression strongly decreases in ciCLMs after 7 and 14 days of differentiation compared to the controls; , 2-way ANOVA test, . (d) Examples of FACS analysis on ciCLMs confirmed a reduced number of Thy1 (CD90) + cells after 7 days of differentiation.